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Custom MHC-peptides Tetramer Service

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Custom MHC-peptides Tetramer Service

Creative Peptides is offering MHC tetramer synthesis with a set of peptide sequences customized by customers now. Here is a list of available alleles that Creative Peptides can provide. Currently two choices of fluorochrome labeling are available based on customer’s request: phycoerythrin(PE) or allophycocyanin(APC). Creative Peptides also provides clinical grade tetramer for our clients. These highly purified products can be used in immunotherapy trials. For more information please contact our technical team at .

Background of MHC Tetramer

Due to the low affinity of the MHC molecule (Major Histocompatibility Complex) to the specific T cell, the study of antigen-specific T cell response is progressing slowly. Until the development of a MHC tetramer assay or a MHC tetramer technology by Stanford University School of Medicine, new doors of possibility were opened up in this filed of research. With the onset of MHC tetramer technology, it comes with a wealth of new understanding of antigen specific T cells.
MHC tetramer technology is an innovative method of directly staining freshly isolated antigen-specific T-cells, which has greatly enhanced our ability to assess cellular dynamics during an immune response. Joining multiple copies of the MHC/antigen complex into a single probe, tetramer technology, resolves the difficulties presented by the low affinity of MHC molecule to the specific T cell.

What Is an MHC Tetramer and How Is the MHC Tetramer Made?

MHC tetramers is a complex of 4 MHC molecules associated with loaded peptide, which is bound to a fluorescently labeled streptavidin molecule. Monomeric MHC molecule is loaded with its cognate peptide, and biotin is linked to the complex at a site far removed from the T-cell recognition site.(Constantin, Bonney, Altman, & Strickland, 2002)

The monomeric MHC-peptide complexes bind to streptavidin, which further tightly binds biotin at 4 separate sites, thus making a tetrameric complex, the MHC tetramer.

MHC tetramer is an innovative product of directly staining freshly isolated antigen-specific T-cells. The isolated cells can be cultured, multiplied and stored for further study use. In the production of tetramers, the MHC heavy chain protein, the light chain protein and the epitopic peptide are refolded into MHC complex in vitro. The successfully refolded monomers are purified by gel-filtration and then biotinylated and stored at –80 °. The biotinylated monomers are bound to fluorescent- labeled streptavidin, generally APC or PE before use. Streptavidin has 4 available binding sites for biotin, thus making a tetramer of MHC molecules with bound peptides. Details of the protocols can be found at NIH Tetramer Core Facility's website.

Note: Custom tetramers may differ significantly in refolding, stability and staining of T cells. It is much dependent on the MHC alleles and the binding affinity of a specific peptide to the MHC molecule. Creative Peptides recommends testing of every new batch of MHC tetramers with positive and negative control cells by titration of the tetramer reagent. Tetramers should be stored at 4°C and retested after long-term storage.

References:
1.Constantin, C. M., Bonney, E. E., Altman, J. D., & Strickland, O. L. (2002). Major histocompatibility complex (MHC) tetramer technology: an evaluation. Biol Res Nurs, 4(2), 115-127. doi:10.1177/1099800402238332.
2.Davis, M. M., Altman, J. D., & Newell, E. W. (2011). Interrogating the repertoire: broadening the scope of peptide-MHC multimer analysis. Nat Rev Immunol, 11(8), 551-558. doi:10.1038/nri3020.

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